Isolation and biological characterization of chicken amnion epithelial cells

Abstract

Amniotic epithelial cells (AECs) express Oct4, Nanog and Sox-2, which are necessary for maintaining the undifferentiated state of pluripotent stem cells. AECs additionally express CK19, which is a specific marker of epithelial cells, both in vivo and in vitro. In this research, we investigated the biological characteristics and potential for cell therapy of AECs from 6-day-old chicken embryos. We induced the AECs to differentiate into pancreatic islet-like cells (endoderm), adipocytes and osteoblasts (mesoderm) and neural-like cells (ectoderm), and used immunofluorescence and RT-PCR to detect the expression of AECs specific markers. To assess the differentiation capacity of AECs, passage 3 cells were induced to differentiate into adipocytes, osteoblasts, pancreatic islet-like cells and neural-like cells. The AEC markers, Oct-4, Nanog, Sox-2 and CK19, were all positively expressed. Cloning efficiency decreased with increasing passage number. Passage 3 AECs were successfully induced to differentiate into pancreatic islet-like cells, osteoblasts, adipocytes, and neural-like cells. These results suggested that AECs isolated from chicken embryos exhibited the characteristics of the multipotent stem cells. AECs may therefore be ideal candidates for cellular transplantation therapy and tissue engineering.

Differential developmental competence and gene expression patterns in buffalo (Bubalus bubalis) nuclear transfer embryos reconstructed with fetal fibroblasts and amnion mesenchymal stem cells

“…It is a well-known fact that mixed population of cells cultured from amniotic membrane in many species such as human, ovine, chicken, rat, equine and canine also displayed heterogeneous cell population (Marcus et al 2008;Mauro et al 2010;Bacenkova et al 2011;Coli et al 2011;Gao et al 2012;Park et al 2012). In agreement with these reports, it was observed that cultured cells exhibited both spindleshaped fibroblasts and polygonal shaped epithelial cell-like morphology in vitro.…”

“…These results indicate that these cells were in an undifferentiated state. Expression of these pluripotency marker genes has previously been reported in amnion-derived cells of various mammalian species including rat (Marcus et al 2008), canine (Park et al 2012) and chicken (Gao et al 2012). In addition, reverse transcriptase PCR results indicated that AMSCs expressed the mesenchymal stem cells markers CD44 and CD29, but not haematopoietic surface markers such as CD34 as reported earlier in bovine .…”

“…Recently, Yamahara et al (2014) reported that mesenchymal stem cells isolated from human amniotic tissue have an angiogenesis potential. In animals, amnion-derived stem cells have potential of differentiation into multilineage mesenchymal cell types in many species such as bovine , canine (Park et al 2012), feline (Rutigliano et al 2013), chicken (Gao et al 2012), rat (Marcus et al 2008) and equine (Coli et al 2011;Violini et al 2012;Seo et al 2013). Further, the amnion derived MSCs are reported to express certain key pluripotency markers including OCT4 .…”

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“…33 AMSCs are multipotent cells and express transcription factors which are essential for maintaining the self-renewal and undifferentiated state (Table 1). AMSCs are able to give rise into osteoblasts, adipocytes, myocardial cells, 34 neural-like cells, and islet like cells.…”

“…AMSCs are able to give rise into osteoblasts, adipocytes, myocardial cells, 34 neural-like cells, and islet like cells. 33 AMSCs suitable tool for tissue engineering research and autologous cell therapy. [34][35][36] Avian cartilage stem/progenitor cells Avian cartilage-derived stem/progenitor cells (CSPCs) were obtained from the surface zone of articular cartilage from 20days old chicken embryo.…”